Confocal Microscopic Observations on Actin Filament Distribution in Lily Pollen Protoplasts

Actin filaments (F-actin) were localized in the isolated pollen protoplasts of lily using TRITC-phalloidin probe and confocal microscopy. Two kinds of pollen protoplasts were examined: one from pollen grains of non-dehiscent anthers(referred to as ‘nearly mature’ pollen); and the other from pollen grains of just dehiscent anthers(referred to as ‘just mature’ pollen). In the cytoplasm of the pollen protoplasts of the ‘nearly mature’ pollen there was a very well organized actin network made up of thick actin bundles. Two types of bundle connections were seen in the network; namely ‘branch’ connections and 'junction' connections. The ‘branch’ connection (or branching points) was formed due to branching or merging of bundies. The ‘junction’ connection (or 'junction' point) had two or more bundles associated with it. Some of the ‘junction’ points might be actin filament organization: centres. The generative cell in iht pollen protoplasts of the ‘nearly mature’ pollen also contained an actin network. But this network was structurally quite loose and the pundles made up the network were short and thick. In the cytoplasm of the pollen protoplasts of the ‘just mature’ pollen the actin net work was more densely packed. The bundles made up the network were also thinner. The actin network in the generative cell was, however, less densely packed. If the pollen protoplasts from both the ‘nearly mature’ and the 'just mature' pollen grains were transferred from a B5 medium into a Brewbaker and Kwack medium supplemented with sucrose, protoplasts rapidly (i.e. within 2 to 3 hours) developed vacuoles and transvacuolar strand. In these va cuolated protoplasts the vegetative nucleus andthe generative cell became tightly surrounded by a new actin network. In the transvacuolar strands there were numerous actin bundles. The “ends” of some of these bundles appeared to be tightly attached to the protoplast membrane indicating that some kind of structures might be present in the protoplast membrane for actin filament attachment.     

龙牙百合的组织培养

1　植物名称　百合 (Ｌｉｌｉｕｍｂｒｏｗｎｉｉ)品种龙牙百合。2　材料类别　当年收获的百合鳞茎。3　培养条件　 (1 )诱导鳞片分化培养基 :ＭＳ ＮＡＡ0 .5ｍｇ·Ｌ-1(单位下同 ) ＧＡ 2 .0 ;(2 )诱导小鳞片分化的培养基 :ＭＳ 6 ＢＡ1 .0 ＮＡＡ 0 .5 ;(3 )诱导小鳞片形成愈伤组织 :ＭＳ 6 ＢＡ 4.0 2 ,4 Ｄ 4.0 ;(4)促进小鳞茎增重培养基 :ＭＳ 6 ＢＡ1 .0 2 ,4 Ｄ0 .5 ＮＡＡ 0 .2 7%糖 0 .8%琼脂 0 .2 %活性炭 ;(5 )液体培养基 :ＭＳ 6 ＢＡ 1 .0 ＮＡＡ 0 .5。以上培养基除 (4)、(5 )外 ,均含 4%蔗糖、0 .7%琼脂 …     

东方百合鳞片的组织培养

东方百合最佳诱导分化培养基为MS 6-BA 1.0 NAA 0.5 2,4-D 1.0;不定芽增殖最佳培养基为MS 6-BA 1.0 NAA 0.1;最佳生根培养基为MS NAA 0.2.3个品种外部鳞片和中部鳞片的分化能力大于内部鳞片;同一个鳞片上段鳞片的分化能力最差,下段最强;每块鳞片分化不定芽数也是上段最少,下段最多.     

百合组织培养和遗传转化的研究进展

百合是单子叶球茎类观赏花卉,具有食用和药用的作用。本文综述了国内外百合组织培养和遗传转化研究进展。包括营养器官、生殖器官和原生质体再生系统的建立。详细介绍了基因转化的方法,例如农杆菌、基因枪、电激穿孔等。浅析百合外源基因的表达,例如GUS、半夏凝集素基因、几丁质酶基因等。同时,讨论了百合遗传转化中存在的问题,以期为百合基因工程方面的研究提供依据。     

Cytological Studies on Tissue Culture of Pinus cembra

Callus tissue of Swiss stone pine, Pinus cembra L. var. sibirica Loud., has been grown successfully for 9 months through 7 transfers. Excellent initial growth was obtained from hypocotyl segments of 7-day-old seedlings. A complex agar medium was used supplemented with 2 mg/l 2,4-D, 0.05 mg/l kinetin, 1 g/l Edamin and 10% v/v coconut milk. No deterioration of growth was observed on this medium after numerous transfers. The callus tissue did not produce shoots or roots, but showed an ability for cytodifferentiation. Tracheids or tracheid-like structures were formed in every passage, but the tracheids of the primary callus culture differed markedly from those of the subcultures. The tracheids of the primary culture probably already existed in the hypocotyl, whereas those of the subcultures were formed in vitro. The callus tissue was mainly diploid during the period studied, and more than 86% of the mitoses were diploid. A few mitoses with tetraploidy or a higher ploidy also occurred, but the tissue did not show any tendency to polyploidization.     

大型海藻的组织培养及其应用

文章对大型海藻组织培养中愈伤组织和原生质体的获得、除菌效果以及植物生长调节物质的选用的研究进展进行了概述,并对其应用和研究前景作了分析和展望。     

In Vitro Induction of Haploid Plantlets from Unpollinated Young Ovaries of Lily and Its Embryo logical Observations

Unpollinated young ovaries of lily (Lilium davidii var. willmottiae (Wilson) Roffill) were inoculated on modified MS medium and N6 medium. Ovary cultures were incubated at 25–28℃, and illuminated with a fluorescent light of about 800–1200 Lux. Cultured ovaries gradually became thicker and longer after 10 days. The calli (about 6–12 mm in size) were produced after 40 days. The calli were then transferred to the differentiation medium. After 50 days, regeneration plantlets were formed. Embryoids were directly produced from some ovaries, which then developed into plantlets. Observation of chromosome number of regeneration plants shows: 65.71% regeneration plants are haploid plants, 34.29% are diploid. Embryological observation of ovary culture shows that haploid plants are from megaspore tetrad, while diploid plants are probably from nucellus cell.     

单抗免疫斑点法和组织印迹法检测百合无症病毒

应用抗百合无症病毒（Lily symptomless virus,LSV）的单克隆抗体,建立了快速检测田间样品的免疫斑点法（Dot—ELISA）和组织印迹法（Tissue blot-ELISA）体系。LSV单抗稀释2,000倍时,Dot-ELISA中病叶粗汁液可被检出的最大稀释度为1：640。Tissue blot—ELISA中样品一次平切后第1次印迹与Dot—ELISA样品1：40稀释的结果相当,前4次印迹均可获得满意的显色效果。常规Tissue blot-ELISA的灵敏度低于Dot—ELISA,但用丙酮处理点过样的硝酸纤维素膜后,二者的灵敏度相当,且Tissue blot—ELISA操作更简便,适合田间大量样品的快速检测。     

红豆杉组织培养及其产物紫杉醇研究进展

该文对红豆杉组织培养过程中外植体的选择、培养基、培养方法等方面研究进展进行了综述。获取抗癌药物紫杉醇主要途径有:红豆杉组织培养途径、人工种植途径、化学合成及微生物生产途径等。     

光调控在植物组织培养中的应用研究进展

光调控是植物组织培养中一种有效的环境控制技术。该文对近年来国内外有关光调控在植物组织培养中的应用,即光照强度、光周期、光质对组培植物的生长发育、光合作用、愈伤组织诱导及其增殖与分化、器官和体细胞胚发生、生理特性及次生代谢物质等方面的影响研究进展进行了综述,为植物细胞工程提供参考。     

银杏快速繁殖及脱毒技术研究

银杏组织培养的实验研究表明：利用胚萌发时实生苗的茎段及新梢为外植体进行离体培养均能诱导出腋芽,经过多次试验后确定,银杏的启动培养中适宜的培养基为MS NAA0．1mg儿;诱导分化阶段培养基为改良MS 6-BAl．0-5．0mg/L NAA0．1％-0．5mg/L 10％CM(椰乳) 0．1％-0．2％活性碳;继代培养阶段适宜培养基为改良MS 6-BAl．0％-3．0mg/L NAA0．2mg/L 0．1％-0．2％活性碳;生根培养阶段适宜培养基为1／2MS IBAl．0(IBAl．5)mg/L,在诱导分化中附加的天然有机成分(椰乳、活性炭等)起到了关键性作用．通过以上各阶段的培养达到了快速繁殖的目的,在短时间内可提供大量银杏幼苗。     

Cytological Observations of Microsporogenesis and Pollen Development in Wheat in Vivo

Cytological observations of microsporogenesis and pollen development in vivo in wheat were carried on by means of phase contrast optics, which could avoid the distortions resulted from materials difficult to be fixed. The cytological changes were observed as follows: 1. Just before first mitosis of pollen, many strands of cytoplasm arose from one side of the nucleus facing the aperture, and moved swingingly toward the aperture. And then these strands of cytoplasm combined into one mass and protruded in the large vacuole. 2. There was a alteration in the direction of the spindle axis from obliquity to parallelism in anaphase. 3. Forming course of the wall between generative and vegetative cell. 4. Dynamic course of the disintegration of wall between vegetative and generative cell as well as that the generative cell came into the vetetafive cell.     

Cytological Observations on Some Species of Dryopteris and Polystichum from China

Cytological observations on five Dryopteris species and four Polystichum species from China are reported. Most of the materials examined were field-fixed in Emei Shan, Sichuan province, and one of them was in Dali Xian, Yunnan province. Only the count of Dryopteris goeringiana was made from the plant brought from Wuling Shan, Beijing and grown in the garden of our Institute. All the materials were fixed in 1:3 acitic acidalcohol, and stained with acetocamine. All the counts were made at diakinesis or metaphase I of meiosis in SMCs .The results of the observation on them are summarized in table 1. Of these, the chromosome numbers for six species are first report. Five species are sexual: diploid: D. goeringiana, D. porosa, D. subinaequalis, D. yui and P. stenoghyllum. Three spe cies are sexual tetraploid: P. acutidens, P. erosum and P. moupingense. Only D. juxtaposita is an apomictic triploid. The vouchers are preserved in the Herbarium of our Institute (PE). We are indebted to Prof. R. C. Ching and C. R. Fraser-Jenkins for identification of the materials.     

Growth of Leptospira pomona and Its Effect on Various Tissue Culture Systems

Miller, Robert E. (University of Nebraska College of Medicine, Omaha), Norman G. Miller, and Roberta J. White. Growth of Leptospira pomona and its effect on various tissue culture systems. J. Bacteriol. 92:502-509. 1966.-Leptospira pomona strain 3341 was grown in association with primary fetal bovine kidney (PBK) and human embryonic skin-muscle fibroblastic (HE) cells in Eagle's minimal essential medium (MEM) with 5% sheep serum. Growth curves of leptospires in PBK and HE cell cultures showed no substantial increase in growth above that obtained in Eagle's MEM in the absence of tissue culture cells. This suggested that no stimulatory growth factors for leptospires were produced by the tissue cells. Fibroblastic cells of the PBK monolayer showed separation, deterioration, and, finally, complete disintegration. Epithelial-like cells remained unaffected. HE cells showed the same cytopathic effect as PBK fibroblastic cells, indicating that this effect was not limited to PBK fibroblastic cells. Warthin-Starry stains of PBK and HE cell monolayers showed masses of leptospires adhering to fibroblastic cells, whereas only a few were seen on epithelial-like cells. Large numbers of leptospires on the surface of fibroblastic cells are very likely associated with the cytopathic effect. Dislodgment of leptospires from fibroblastic cells did not increase the total number of spirochetes in the culture. This indicated that leptospiral growth did not occur on the surface of these cells.     

Light and Electron Microsopic Observations on the Pathogenesis of Naegleria fowleri in Mouse Brain and Tissue Culture

Two strains of pathogenic Naegleria were employed to infect mice and monkey kidney (Vero line) cell cultures. Mice were infected intranasally. Moribund mice were sacrificed and their brains processed for light and electron microscopy. The normal architecture of the infected brain was completely destroyed; the olfactory lobes and the cerebral cortex showed the heaviest damage. The inflammatory response was mainly in the form of neutrophil polymorphs (PMN) and was confined to the olfactory lobes and the superficial regions of cerebral cortex. Numerous amebas were seen interspersed with the degenerating neurons, glial processes, and PMN. Most conspicuous were the food vacuoles which contained host tissue in various stages of digestion. Amebas in the brain tissue also produced many micropinocytotic vesicles from the surface of the plasma membrane. These vesicles are interpreted as vehicles of transport of nutritive materials from the host tissue. The infected cell culture showed the characteristic cytopathic effect (CPE). The CPE was chiefly in the form of cell shrinkage, nuclear pycnosis and discontinuity of cell sheet. Amebas were often seen in an intracellular location. The Vero cells produced many fuzzy pinocytotic vesicles at these loci where the ameba plasma membrane and Vero cell membrane were in close apposition; the probable significance of this is discussed. Most impressive, however, were the pseudopodial formation and capturing of the host material which indicated the great phagocytic activity of the amebas. This was confirmed further by the presence of large numbers of food vacuoles containing host material in various stages of digestion. These observations show that the amebas invade and destroy the brain tissue by active phagocytosis.     

百合未授粉子房离体培养胚胎形成及植株再生

未受精子房离体培养是诱导雌核产生单倍体的技术之一。以1个野生种和3个杂种系共7个百合(Lilium)基因型为实验材料, 探讨了基础培养基、花蕾取样时期和外源激素等因素对百合未授粉子房离体培养胚胎形成的影响。结果表明, CBM、MS和BDS三种基础培养基均可诱导百合未授粉子房胚胎形成, 但以BDS培养基诱导效果最佳; 开花前1天的花蕾较适于百合未授粉子房离体培养; 2 mg·L^-1 2,4-D + 2 mg·L^-1 6-BA和2 mg·L^-1 2,4-D + 2 mg·L^-1 KT两种外源激素配方均适用于百合未授粉子房离体培养诱导胚胎形成。在培养过程中, 大多数胚性胚珠中只含有1个胚胎, 位于珠孔端、合子端或极核处, 少数胚性胚珠中含有双胚胎。通过百合未授粉子房离体培养, 从5个基因型材料中共获得146株再生植株。采用根尖染色体计数法对其中的62株进行了倍性测定, 其中43株与母体植株染色体倍性不同。     

Cytological Observations on Microsporogenesis and Pollen Development of Regenerated Stamen in Hyacinth

The present study consists of the cytological observations on the process of microsporogenesis and pollen development in the regenerated stamen of hyacinth; and a comparative study of the cytological changes in stamens of both regenerated and produced under natural condition. Results showed that the cytological changes of microsporogenesis and the pollen deveLopment in the regenerated stamen of the hyacinth were basically normal. But in the stage of the mature pollen there was an obvious cytological difference between both stamens in vitro and in nature. The mature pollen of the regenerated stamen consisted of three cells: one vegetative cell and two sperms, while mature pollen grain under natural condition was made up of two cells: one vegetative cell and one generative cell. This difference mainly resulted from different time and place of the generative cell division. The reason resulting in the differences and their influence on sperms were discussed.